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Large-scale translocation reversal within the thylakoid Tat system in vivo
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UNSPECIFIED (2005) Large-scale translocation reversal within the thylakoid Tat system in vivo. JOURNAL OF CELL BIOLOGY, 171 (2). pp. 281-289. ISSN 0021-9525.
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Abstract
In vitro import assays have shown that the thylakoid twin- arginine translocase (Tat) system transports folded proteins in a unidirectional manner. Here, we expressed a natural substrate, pre-23K, and a 23K presequence green fluorescent protein (GFP) chimera in vivo in tobacco protoplasts. Both are imported into chloroplasts, targeted to the thylakoids, and processed to the mature size by the lumen-facing processing peptidase. However, the vast majority of mature GFP and about half of the 23K are then returned to the stroma. Mutations in the twin-arginine motif block thylakoid targeting and maturation, confirming an involvement of the Tat apparatus. Mutation of the processing site yields membrane-associated intermediate-size protein in vivo, indicating a delayed reversal of translocation to the stroma and suggesting a longer lived interaction with the Tat machinery. We conclude that, in vivo, the Tat system can reject substrates at a late stage in translocation and on a very large scale, indicating the influence of factors that are absent in reconstitution assays.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QH Natural history > QH301 Biology | ||||
Journal or Publication Title: | JOURNAL OF CELL BIOLOGY | ||||
Publisher: | ROCKEFELLER UNIV PRESS | ||||
ISSN: | 0021-9525 | ||||
Official Date: | 24 October 2005 | ||||
Dates: |
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Volume: | 171 | ||||
Number: | 2 | ||||
Number of Pages: | 9 | ||||
Page Range: | pp. 281-289 | ||||
Publication Status: | Published |
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